Evoked Potentials ; Social Isolation ; Attention ; Humans

Objective To make the bioinformatics analysis of Ureaplasma parvum UP3-RS02445 and prepare monoclonal antibody (mAb) against UP3-RS02445. Methods The biological characteristics of UP3-RS02445 protein were predicted by bioinformatics software. The UP3-RS02445 prokaryotic expression plasmid was constructed and the corresponding protein expression was induced by isopropyl-β-D-thiogalactoside (IPTG). Thus the expressed protein was used as immunogen to immunize female BALB/c mice. Hybridoma cell technology was used to prepare the monoclonal antibody against UP3-RS02445. The specificity and titer of monoclonal antibody were detected by Western blot and ELISA respectively. The subclass of heavy chain and subtype of light chain were identified by monoclonal antibody subtype identification test strip. Results Bioinformatics analysis showed that UP3-RS02445 protein was composed of 201 amino acids, without transmembrane domain and signal peptide, and belongs to non-secretory proteins. The recombinant prokaryotic plasmid of UP3-RS02445 was successfully constructed and the recombinant protein could be induced in large amount. After cell fusion, two hybridoma cells (A1H5 and A4E2) secreting UP3-RS02445 mAb were screened by ELISA and Western blot. The results of ELISA showed that the titers of monoclonal antibodies were 1:2560. Western blot and Immunofluorescence technique both indicated that the antibodies could bind specifically to the UP3-RS02445 protein. The heavy chain and light chain of the two mAbs were IgG1 and kappa subtypes respectively. Conclusion We prepared the UP3-RS02445 monoclonal antibodies with well specificity and high titer which might lay foundations for the subsequent development of UP diagnostic reagents and the functional study of protein.
Antibodies, Monoclonal/immunology* ; Animals ; Mice, Inbred BALB C ; Female ; Computational Biology/methods* ; Mice ; Ureaplasma urealyticum/genetics* ; Bacterial Proteins/genetics* ; Antibody Specificity ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology*

Objective:To investigate the relationship between the expression of CD103 +CD8 +T cells in locally advanced oral squamous cell carcinoma (LA-OSCC), and the response to neoadjuvant chemoimmunotherapy (NACI). Methods:Thirty LA-OSCC patients from the Department of Oral and Maxillofacial Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, who underwent NACI from June 2020 to December 2022 were analyzed, including 16 responders and 14 non-responders. Using multiple immunofluorescence technique to stain sections of patients to verify the correlation between the expression of CD103 +CD8 +T cells and the efficacy of NACI. CD103 +CD8 +T cell density was counted using Inform and HALO software. The Spearman correlation coefficient in rank correlation is used to describe the correlation between CD103 +CD8 +T cell and neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-lymphocyte ratio (PLR), systemic immune inflammation index (SII) It′s effectiveness as a predictive marker to NACI was analyzed by receiver operator characteristic (ROC) curve analysis and decision curve analysis (DCA). Two-tailed t-test or Mann-Whitney U-test was used to compare data between two groups, and one-way ANOVA was used to compare data between multiple groups. SPSS 22.0 and GraphPad prism 9.0 software were used for statistical analysis and plotting of relevant statistical graphs such as histograms. P<0.05 was considered a statistically significant difference. Results:The density of CD103 +CD8 +T cells has expanded in advanced OSCC patients who are responsive to NACI. The CD103 +CD8 +T cell densities in the responsive and nonresponsive groups were 118.30(41.92, 197.80) pcs/mm 2 and 21.63(4.91, 71.92) pcs/mm 2 respectively, with statistically significant differences( U=52.00, P=0.012). CD103 +CD8 +T cell abundance was negatively correlated with NLR, dNLR, PLR, and SII ( P<0.05). ROC curve analysis showed that the AUC for predicting efficacy of NLR, dNLR, PLR, and SII were 0.781 ( P=0.009, 95% CI: 0.5715-0.9910), 0.671 ( P=0.105, 95% CI: 0.467-0.881), 0.679 ( P=0.020 95% CI: 0.549-0.951), 0.750 ( P=0.096, 95% CI: 0.461-0.896), respectively. The AUC for CD103 +CD8 +T cells alone was 0.861 ( P=0.013, 95% CI: 0.585-0.950), and the AUC of combining CD103 +CD8 +T cells with NLR was 0.896 ( P=0.025, 95% CI: 0.454-0.938). Conclusions:The density of CD103 +CD8 +T cells is expanded in advanced OSCC patients who are responsive to NACI. CD103 +CD8 +T cells positively predict favorable responses as a strong indicator to NACI in advanced OSCC patients. Co-interpretation of CD103 +CD8 +T cells and NLR value enhances the predictive accuracy of NACI in advanced OSCC patients.

BACKGROUND: Angiogenesis is described as a complex process in which new microvessels sprout from endothelial cells of existing vasculature. This study aimed to determine whether long non-coding RNA (lncRNA) H19 induced the angiogenesis of gastric cancer (GC) and its possible mechanism. METHODS: Gene expression level was determined by quantitative real-time polymerase chain reaction and western blotting. Cell counting kit-8, transwell, 5-Ethynyl-2'-deoxyuridine (EdU), colony formation assay, and human umbilical vein endothelial cells (HUVECs) angiogenesis assay as well as Matrigel plug assay were conducted to study the proliferation, migration, and angiogenesis of GC in vitro and in vivo . The binding protein of H19 was found by RNA pull-down and RNA Immunoprecipitation (RIP). High-throughput sequencing was performed and next Gene Ontology (GO) as well as Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was conducted to analyze the genes that are under H19 regulation. Methylated RIP (me-RIP) assay was used to investigate the sites and abundance among target mRNA. The transcription factor acted as upstream of H19 was determined through chromatin immunoprecipitation (ChIP) and luciferase assay. RESULTS: In this study, we found that hypoxia-induced factor (HIF)-1α could bind to the promoter region of H19, leading to H19 overexpression. High expression of H19 was correlated with angiogenesis in GC, and H19 knocking down could inhibit cell proliferation, migration and angiogenesis. Mechanistically, the oncogenic role of H19 was achieved by binding with the N 6 -methyladenosine (m 6 A) reader YTH domain-containing family protein 1 (YTHDF1), which could recognize the m 6 A site on the 3'-untransated regions (3'-UTR) of scavenger receptor class B member 1 (SCARB1) mRNA, resulting in over-translation of SCARB1 and thus promoting the proliferation, migration, and angiogenesis of GC cells. CONCLUSION HIF-1α induced overexpression of H19 via binding with the promoter of H19, and H19 promoted GC cells proliferation, migration and angiogenesis through YTHDF1/SCARB1, which might be a beneficial target for antiangiogenic therapy for GC.
Humans ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Endothelial Cells/metabolism* ; Gene Expression Regulation ; Gene Expression Regulation, Neoplastic/genetics* ; Hypoxia ; MicroRNAs/genetics* ; RNA ; RNA, Long Noncoding/metabolism* ; RNA-Binding Proteins/metabolism* ; Scavenger Receptors, Class B/metabolism* ; Stomach Neoplasms/genetics*

Chlamydia, a gram-negative obligate intracellular pathogen, is a major cause of human reproductive tract, eye and respiratory tract infections. It replicates in a special membrane-binding chamber called inclusion and survives in the host′s hostile intracellular environment through secreting effectors, but requires host-derived lipids to grow and develop in the cells. Emerging evidences suggest that Chlamydia has evolved a variety of strategies to meet its lipid needs by interacting with host cell compartments and redirecting the transport pathway to its intracellular niche. This paper briefly described the pathway for obtaining host lipids and the mechanism of lipid metabolic during Chlamydia infection.

Tocilizumab has been reported to attenuate the "cytokine storm" in COVID-19 patients. We attempted to verify the effectiveness and safety of tocilizumab therapy in COVID-19 and identify patients most likely to benefit from this treatment. We conducted a randomized, controlled, open-label multicenter trial among COVID-19 patients. The patients were randomly assigned in a 1:1 ratio to receive either tocilizumab in addition to standard care or standard care alone. The cure rate, changes of oxygen saturation and interference, and inflammation biomarkers were observed. Thirty-three patients were randomized to the tocilizumab group, and 32 patients to the control group. The cure rate in the tocilizumab group was higher than that in the control group, but the difference was not statistically significant (94.12% vs. 87.10%, rate difference 95% CI-7.19%-21.23%, P = 0.4133). The improvement in hypoxia for the tocilizumab group was higher from day 4 onward and statistically significant from day 12 (P = 0.0359). In moderate disease patients with bilateral pulmonary lesions, the hypoxia ameliorated earlier after tocilizumab treatment, and less patients (1/12, 8.33%) needed an increase of inhaled oxygen concentration compared with the controls (4/6, 66.67%; rate difference 95% CI-99.17% to-17.50%, P = 0.0217). No severe adverse events occurred. More mild temporary adverse events were recorded in tocilizumab recipients (20/34, 58.82%) than the controls (4/31, 12.90%). Tocilizumab can improve hypoxia without unacceptable side effect profile and significant influences on the time virus load becomes negative. For patients with bilateral pulmonary lesions and elevated IL-6 levels, tocilizumab could be recommended to improve outcome.
Antibodies, Monoclonal, Humanized ; COVID-19/drug therapy* ; Humans ; SARS-CoV-2 ; Treatment Outcome

Objective:To investigate the expression and phosphorylation level change of adenosine monophosphate activated protein kinase (AMPK) in skeletal muscle of severely scald rats and its roles in skeletal muscle atrophy in severely scalded rats.Methods:The experimental research method was applied. Totally 100 6-week-old male Wistar rats were divided into sham injury group and scald group according to the random number table, with 50 rats in each group. After weighing the body weight, rats in scald group were inflicted with full-thickness scald of 30% total body surface area on the back, and rats in sham injury group were simulated with scald. At 6 h and on 1, 3, 5, and 7 d post injury, 10 rats in each group were taken to measure their body weights and weights of extensor digitorum longus and soleus muscle. At 6 h and on 1, 3, 5, and 7 d post injury, the tibialis anterior muscles were collected, the mRNA expressions of muscle atrophy F-box protein (MAFbx) and muscle-specific RING finger protein 1 (MuRF1) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction; the content of adenosine monophosphate (AMP), adenosine diphosphate, and adenosine triphosphate (ATP) were detected by high performance liquid chromatography, and AMP/ATP ratio and energy charge were calculated; the protein expressions of AMPK-α and phosphorylated AMPK-α (p-AMPK-α) were detected by Western blotting, and the p-AMPK-α/AMPK-α ratio was calculated, with sample number of 4 in each time point of each group. Data were statistically analyzed with analysis of variance for factorial design and least significant difference test.Results:The body weights of rats in 2 groups before injury and at each time point post injury were close ( P>0.05). At 6 h post injury, the weight of extensor digitorum longus of rats in scald group was (0.107±0.007) g, which was significantly heavier than (0.086±0.0607) g of sham injury group ( P<0.01). On 3 d post injury, the weight of extensor digitorum longus of rats in scald group was (0.083±0.016) g, which was significantly lighter than (0.102±0.005) g of sham injury group ( P<0.01). The weight of soleus of rats in 2 groups were close at each time point post injury ( P>0.05). Compared with those of sham injury group, the mRNA expression of MAFbx in tibialis anterior muscle of rats in scald group was significantly up-regulated at 6 h post injury ( P<0.01), and the mRNA expressions of MuRF1 in tibial anterior muscle of rats in scald group were significantly up-regulated at 6 h and on 1 d post injury ( P<0.01). At 6 h and on 7 d post injury, compared with those of false injury group, the AMP/ATP ratios of the tibial anterior muscle of rats in scald group were significantly increased ( P<0.05 or P<0.01), and energy charges of the tibial anterior muscle of rats in scald group were significantly decreased ( P<0.01). At each time point post injury, the protein expressions of AMPK-α of the tibial anterior muscle of rats in 2 groups were close ( P>0.05). The p-AMPK-α/AMPK-α ratios of the tibial anterior muscle of rats in scald group at 6 h and on 7 d post injury were significantly higher than those in sham injury group ( P<0.05 or P<0.01). Conclusions:The decrease in energy charge and increase in AMP/ATP ratio of skeletal muscle of rats after severe scald activate AMPK. The activation of AMPK in the early stage of injury is consistent with the up-regulation of MAFbx and MuRF1 expressions and down-regulation of skeletal muscle weight. The above-mentioned changes may be one of the molecular mechanisms of skeletal muscle atrophy in rats with severe scald

Objective To investigate the clinical effect of monosialotetrahexosyl ganglioside in the treatment of neonatal intracranial hemorrhage.Methods From January 2016 to December 2018,142 neonates with intracranial hemorrhage admitted to the Maternal and Child Health Hospital of Zhoushan were randomly divided into observation group (71 cases) and control group (71 cases) according to the digital table.The control group was treated with routine treatment,while the observation group was treated with ganglioside needle on the basis of the control group.Both two groups were treated for 14 days.The therapeutic effects,muscle tone recovery time,reflex recovery time and consciousness recovery time were compared.The changes of neurobehavioral assessment score (NABA score),TNF-αt,IL-1β,MMP-2,T IMP-1 and NSE levels before and after treatment were compared.Results The total effective rate of the observation group (92.96％) was higher than that of the control group (77.47％) (x2 =6.762,P ＜ 0.05).The recovery time of muscle tension,reflex and consciousness in the observation group [(7.68 ± 1.29) d,(6.83 ± 1.20) d and (8.34 ± 1.54) d] were shorter than those in the control group [(10.25 ± 2.31) d,(9.17 ±1.86) d and (10.53 ± 1.08) d] (t =8.185,8.908,9.811,all P ＜ 0.05).After treatment,the NABA score of the observation group [(40.37 ± 0.65) points] was higher than that of the control group [(37.16 ± 0.93) points] (t =23.838,P ＜ 0.05).The serum levels of TNF-α [(26.37 ± 4.25) pg/L],IL-1β [(16.74 ± 3.24) ng/L],MMP-2 [(78.39 ± 16.57)g/L],TIMP-1 [(179.32 ± 17.65) ng/mL] and NSE [(13.52 ± 2.19) g/L] in the observation group were lower than those in the control group [(53.21 ± 7.39) pg/L,(28.93 ± 5.64) ng/L,(97.42 ±12.63) g/L,(238.63 ± 28) ng/mL and (21.43 ± 2.89) μg/L] (t =26.529,15.792,7.696,14.938,1 8.381,all P ＜ 0.05).Conclusion Ganglioside has good therapeutic effect on neonatal intracranial hemorrhage.It can reduce the serum levels of TNF-α,IL-1β,MMP-2,TIMP-1 and NSE,and improve the neurobehavioral function of neonates.

Objective To investigate the relationship between serum markers β amyloid (Aβ), tau and thyroid hormone levels and post-stroke cognitive impairment (PSCI) in the acute phase of cerebral infarction. Methods A total of 214 patients with acute cerebral infarction were enrolled. The baseline data and serological indicators were collected and the cognitive function of patients was evaluated. All patients were divided into cognitive impairment group and normal group based on follow-up results. The differences of Aβ1-42, tau protein and thyroxine levels between the two groups and their relationship with disease progression were analyzed. The Cox regression analysis and ROC curve were used to compare the above parameters to predict the development of PSCI. Results The total protein level of Tau (210.6 ±98.9 pg/mL) was higher and Aβ1-42 (426.1 ±123.5 pg/mL) and triiodothyronine (T3) (1.43 ±0.57 nmol/L), free thyroxine (FT4) (13.15±2.23 pmol/L) was significantly lower in the cognitive impairment group than in the normal group (P<0.05). Tau protein (r=-0.457), Aβ1-42 (r=0.348), T3 (r=0.211), and FT4 (r=0.306) were all associated with disease progression (P<0.05). Cox regression analysis showed that Aβ1-42 and T3 were important influencing factors in the occurrence of PSCI. The area under the curve of Aβ1-42 combined with T3 was 0.841. The specificity and the sensitivity were 74.8% and 85.3%, respectively, with a diagnostic cutoff value of 0.572. Conclusion Aβ1-42 and T3 levels in the acute phase of cerebral infarction may predict the progression of PSCI.

Objective To investigate the efficacy of leflunomide combined with prednisone in the induction therapy of proliferative lupus nephritis (LN).Methods A prospective,multicenter,randomized controlled clinical trial was conducted in patients with biopsy-proved proliferative lupus nephritis recruited from 15 renal centers from 2013 to 2015.Patients were randomized to two groups.Oral leflunomide or intravenous cyclophosphamide was given to patients in each group.Both groups received a tapering course of oral prednisone therapy.All patients were followed up for 24 weeks.The blood biochemistry,urine index,clinical curative effect and adverse reaction were recorded and analyzed statistically.Results A total of 100 patients were enrolled in this clinical trial,including 48 patients in leflunomide group and 52 patients in cyclophosphamide group.After 24 weeks,the overall response rate was 79％ (95％ CI 67％-90％) in the leflunomide group and 69％ (95％ CI 56％-82％) in the cyclophosphamide group.23％ (95％CI 11％-35％) of patients in leflunomide group showed complete remission compared with 27％ (95％CI 24％-30％) in cyclophosphamide group (P=0.35).The levels of 24-hr urine protein excretion,SLEDAI and anti-dsDNA antibody titers were decreased in patients treated with leflunomide group after 24-weeks treatment.And the levels of serum albumin and complement 3 after treatment were significantly higher compared with these before treatment.There was also no significant difference in changes of 24-hr urine protein excretion,SLEDAI score,anti-dsDNA antibody titers,serum albumin and complement C3 levels after treatment between two groups.Incidence of adverse events did not differ between the leflunomide and cyclophosphamide group.Conclusions Leflunomide combined with prednisone showed same efficacy compared with cyclophosphamide as induction therapy for lupus nephritis.Leflunomide might be an useful medicine in the induction therapy of lupus nephritis.